Summary: Single-cell sequencing (SCS) is a disruptive new technology that offers an unprecedented ability to interrogate the genomics (DNA, RNA) of single cells to understand the complex biologies of normal and diseased tissues. SCS is not only important for basic research, but also represents a next generation of diagnostic approaches?particularly for cancer. While multiple SCS platforms have been marketed or are under development, all of these rely on high-quality preparations of single, viable cells in a fluid suspension to enter their respective workflows. This is not a trivial undertaking, as enzymatically digested normal or diseased tissues contain dead or dying cells and cell aggregates that may confound SCS. Current cell preparation methods for SCS, such as fluorescence-activated cell-sorting (FACS) or dead-cell removal kits, are suboptimal in that they 1) often require a significant investment of time, money and/or training; 2) require multiple protocols; and/or 3) result in a poor yield of viable cells, which is particularly problematic for scarce clinical specimens. The SynderBio R&D team has developed a proprietary approach for separating cell types based on their differential responses to brief pulses of fluid shear stress (FSS). Our preliminary data supports our hypothesis that we can use this technology to rapidly (<15 min) eliminate dead/dying cells and dissociate cell aggregates from cell suspensions. Our Specific Aims and Milestones for this Phase I SBIR feasibility project are as follows: Specific Aim 1: Calibrate the performance characteristics of SynderBio's FSS technology for SCS. Milestone: Achieve ?95% viable, single-cell preparation with ?80% yield. Specific Aim 2: Compare SynderBio's FSS technology to existing cell-preparation methods. Milestones: Achieve significantly better performance characteristics (<30min, ?95% viable single cells, ?80% yield) with purity comparable to that of FACS or dead-cell removal kits from competitors. Phase II Aims will then be to 1) transform SynderBio's current pre-prototype device into a commercial-ready product; 2) distribute a limited number of these devices to thought leaders and potential strategic partners in the field; and 3) expand the side-by-side comparison with competing technologies to other relevant applications in the field. Phase II work will be focused on successful prototyping and in-depth testing designed to obtain the data required by potential Phase III industry collaborators/investors to engage in the next steps required for Phase III commercialization. The commercial opportunity for SynderBio's product is significant. The global cell-isolation/separation market alone is expected to reach $7.89B in 2021 at a CAGR of 17.2%. SynderBio's technology is also applicable to cell preparation for other purposes, such as FACS. Thus, we expect the successful development of SynderBio's technology to have a major positive impact on human health and to address significant commercial opportunities related to the use of SCS for understanding and differentiating normal and diseased tissue biology.